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文献和实验DpnI mediated site-directed Mutagenesis
the amplification to work efficiently with pfu take a bit of fiddling, amplification with Ex taq appears very robust for large plasmids. REFERENCES Fisher, C. L., and Pei, G. K. (1997). Modification of a PCR -based site-directed mutagenesis method
Site-Directed Mutagenesis In Vitro by Megaprimer PCR
Among the various mutagenesis procedures based on polymerase chain reaction (PCR), the “megaprimer” method appears to be the simplest and most versatile. The method utilizes three oligonucleotide primers and two rounds of PCR performed on a DNA
Site-directed Mutagenesis using PCR
product, and (vi) efficient intramolecular ligation in the presence of T4 DNA ligase.Protocol. PCR-based Site Directed MutagenesisPlasmid template DNA (approximately 0.5 pmole) is added to a PCR cocktail containing, in 25 ul of 1x mutagenesis buffer: (20
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